The overall objective of this proposal is to complete the development of reliable and reproducible systems and methods for the large-scale culture of large-egged octopods, thus allowing the scheduled production of these highly advanced invertebrates for biomedical research in the United States. In Europe and Japan, Octopus has been and continues to be a well established model for biological experimentation because of its well developed organ systems, particularly the nervous system which is highly organized and extensively studied. Moreover, octopuses are good research animals because their behavior is similar to vertebrates, they are adaptable to reward and punishment training paradigms, they have short- and long-term memory stores, they may be narcotized for long periods and they withstand operative procedures well. Little work has been done on Octopus in the U.S. because the native species do not occur in large, easily harvested populations, but laboratory culture has been shown to be a feasible means of providing them. The specific aims of this proposal include culturing at least 100 Octopus bimaculoides to a size of 250g in each of two culture generations. Brood stock management procedures will be developed to minimize inbreeding depression, to enhance hatchling fitness and to increase the number and quality of eggs produced. The temporal availability of eggs and octopuses will be expanded through manipulation of temperature and light periodicity, while the array of live and dead food organisms that octopuses can be reared on at inland locations will be enlarged by evaluating diets of marine polychaete worms and freshwater organisms. Methods to increase the culture efficiency of recirculating seawater systems will be evaluated to develop detailed guidelines for water management and for the design, construction and maintenance of small- and large-scale octopus culture systems. Reliable methods will be developed for transporting octopuses long distances, and cost estimates will be calculated for culturing octopuses for research. The octopuses will be utilized in a wide range of experimental applications to assess their specific usefulness in research. By solving the remaining problems of octopus culture and by promoting the use of octopuses among biomedical researchers in the U.S., an important invertebrate species will be added to the repetoire of animals available for investigations of basic biological principles.